Open AccessSmall Molecule Mesengenic Induction of Human Induced Pluripotent Stem Cells to Generate Mesenchymal Stem/Stromal Cells
Author(s) -
Chen Yen Shun,
Pelekanos Rebecca A.,
Ellis Rebecca L.,
Horne Rachel,
Wolvetang Ernst J.,
Fisk Nicholas M.
Publication year - 2012
Publication title -
stem cells translational medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.781
H-Index - 71
eISSN - 2157-6580
pISSN - 2157-6564
DOI - 10.5966/sctm.2011-0022
Subject(s) - mesenchymal stem cell , induced pluripotent stem cell , embryoid body , microbiology and biotechnology , biology , embryonic stem cell , stem cell , cellular differentiation , regenerative medicine , reprogramming , cell , genetics , gene
The translational potential of mesenchymal stem/stromal cells (MSCs) is limited by their rarity in somatic organs, heterogeneity, and need for harvest by invasive procedures. Induced pluripotent stem cells (iPSCs) could be an advantageous source of MSCs, but attempts to derive MSCs from pluripotent cells have required cumbersome or untranslatable techniques, such as coculture, physical manipulation, sorting, or viral transduction. We devised a single‐step method to direct mesengenic differentiation of human embryonic stem cells (ESCs) and iPSCs using a small molecule inhibitor. First, epithelial‐like monolayer cells were generated by culturing ESCs/iPSCs in serum‐free medium containing the transforming growth factor‐β pathway inhibitor SB431542. After 10 days, iPSCs showed upregulation of mesodermal genes ( MSX2, NCAM, HOXA2 ) and downregulation of pluripotency genes ( OCT4, LEFTY1/2 ). Differentiation was then completed by transferring cells into conventional MSC medium. The resultant development of MSC‐like morphology was associated with increased expression of genes, reflecting epithelial‐to‐mesenchymal transition. Both ESC‐ and iPSC‐derived MSCs exhibited a typical MSC immunophenotype, expressed high levels of vimentin and N‐cadherin, and lacked expression of pluripotency markers at the protein level. Robust osteogenic and chondrogenic differentiation was induced in vitro in ES‐MSCs and iPS‐MSCs, whereas adipogenic differentiation was limited, as reported for primitive fetal MSCs and ES‐MSCs derived by other methods. We conclude that treatment with SB431542 in two‐dimensional cultures followed by culture‐induced epithelial‐to‐mesenchymal transition leads to rapid and uniform MSC conversion of human pluripotent cells without the need for embryoid body formation or feeder cell coculture, providing a robust, clinically applicable, and efficient system for generating MSCs from human iPSCs.